I am planning to produce TEV protease in my lab because I will need a lot of the protein in order to cleave the fusion tag off the recombinant proteins which I will be producing in E. coli. Do suggest a good protocol/paper.
Thanks.
Hi Renuk,
cannot send you an article but can share details. We were producing TEV as a fusion with MBP. There was tev site between MBP and TEV so the fusion would be cleaved after being successfully expressed. There was also a His6 tag on TEV and we used it to purify on Ni-agarose, batch method. Avoid high salts, keep pH at 8.0, use TCEP or DTT to constantly maintain a reducing environment. As a Cys protease, it has to be maintained in the reducing environment.
Here is a useful overview of different versions of TEV with corresponding references. There are some derivatives which are more stable (increases yields) and more active. I hope it helps.
https://mcl1.ncifcrf.gov/waugh_tech/faq/tev.pdf
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